Ambiguities and completeness of SAS data analysis: Investigations of apoferritin by SAXS/SANS EID and SEC-SAXS methods

D.V. Zabelskii, A.V. Vlasov, Yu L. Ryzhykau, T.N. Murugova, M. Brennich, D.V. Soloviov, O.I. Ivankov, V.I. Borshchevskiy, A.V. Mishin, A.V. Rogachev, A. Round, N.A. Dencher, G. Büldt, V.I. Gordeliy, A.I. Kuklin

Research output: Chapter in Book / Report / Conference proceedingConference contributionAcademicpeer-review

Abstract

The method of small angle scattering (SAS) is widely used in the field of biophysical research of proteins in aqueous solutions. Obtaining low-resolution structure of proteins is still a highly valuable method despite the advances in high-resolution methods such as X-ray diffraction, cryo-EM etc. SAS offers the unique possibility to obtain structural information under conditions close to those of functional assays, i.e. in solution, without different additives, in the mg/mL concentration range. SAS method has a long history, but there are still many uncertainties related to data treatment. We compared 1D SAS profiles of apoferritin obtained by X-ray diffraction (XRD) and SAS methods. It is shown that SAS curves for X-ray diffraction crystallographic structure of apoferritin differ more significantly than it might be expected due to the resolution of the SAS instrument. Extrapolation to infinite dilution (EID) method does not sufficiently exclude dimerization and oligomerization effects and therefore could not guarantee total absence of dimers account in the final SAS curve. In this study, we show that EID SAXS, EID SANS and SEC-SAXS methods give complementary results and when they are used all together, it allows obtaining the most accurate results and high confidence from SAS data analysis of proteins.
Original languageEnglish
Title of host publication3rd International Summer School and Workshop "Complex and Magnetic Soft Matter Systems: Physico-Mechanical Properties and Structure"
EditorsY. Raikher, A. Rogachev, M. Bunoiu, M. Balasoiu
PublisherInstitute of Physics Publishing
Volume994
DOIs
Publication statusPublished - 28 Mar 2018
Externally publishedYes
Event3rd International School and Workshop on Complex and Magnetic Soft Matter Systems: Structure and Physico - Mechanical Properties, CMSMS 2017 - Dubna, Moscow Region, Russian Federation
Duration: 28 Jun 201730 Jun 2017

Publication series

NameJournal of Physics: Conference Series
ISSN (Print)1742-6588
ISSN (Electronic)1742-6596

Conference

Conference3rd International School and Workshop on Complex and Magnetic Soft Matter Systems: Structure and Physico - Mechanical Properties, CMSMS 2017
Country/TerritoryRussian Federation
CityDubna, Moscow Region
Period28/06/1730/06/17

Funding

We acknowledge the European Synchrotron Radiation Facility for providing synchrotron radiation facilities. We thank ESRF structural biology group for their assistance during the experiments. The investigation used the platforms of the Grenoble Instruct Centre (ISBG; UMS 3518 CNRS-CEA-UJF-EMBL). We also thank Russian Science Foundation (grant No. 16-15-00242); FRISBI (grant No. ANR-10-INSB-05-02); GRAL within the Grenoble Partnership for Structural Biology (PSB) (grant No. ANR-10-LABX-49-01); CEA(IBS) and HGF(FZJ) (contract No. CEA(IBS) – HGF(FZJ) STC 5.1 specific agreement).

FundersFunder number
Russian Science Foundation16-15-00242
French Infrastructure for Integrated Structural Biology

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